Detection methods employing HCV core lipid and DNA binding domain monoclonal antibodies

• US 10,345,311 B2
• Filed: 06/22/2016
• Issued: 07/09/2019
• Est. Priority Date: 03/14/2013
• Status: Active Grant

• Current Assignee: Abbott Laboratories Incorporated
• Original Assignee: Abbott Laboratories Incorporated
• Inventors: Ziemann, Robert, Ahlberg, April, Hawksworth, David, Tieman, Bryan, Muerhoff, A. Scott, Marohnic, Christopher, Otis, Kathy
• Primary Examiner(s): Boesen, Agnieszka
• Application Number:
  US15/189,638
• Publication Number:
  US 20170052183A1
• Time in Patent Office: 1,112 Days
• Field of Search: None
• CPC Class Codes: C07K 16/109 Hepatitis C virus; Hepatitis G virus
• CPC Class Codes: C07K 2317/34 Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: G01N 2333/186 Hepatitis C; Hepatitis NANB
• CPC Class Codes: G01N 2469/10 Detection of antigens from microorganism in sample from host
• CPC Class Codes: G01N 2800/52 Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
• CPC Class Codes: G01N 33/5767 non-A, non-B hepatitis

Abstract

The present disclosure provides detection methods employing HCV core lipid binding domain and DNA binding domain monoclonal antibodies. In certain embodiments, the lipid binding domain monoclonal antibody recognizes an epitope in amino acids 141 to 161 of HCV core protein.

Claims

1. A method for the detection of HCV protein in a test sample comprising:

(i) contacting a test sample suspected of containing HCV with a first monoclonal antibody, or antigen-binding portion thereof, which specifically binds to an epitope in the DNA binding domain of an HCV core protein to form a complex between said first monoclonal antibody, or antigen-binding portion thereof, and said HCV core protein located within said test sample;

(ii) contacting said complex formed in step (i) with a second monoclonal antibody comprising a heavy chain variable domain amino acid sequence of SEQ ID NO;

20 and a light chain variable domain amino acid sequence of SEQ ID NO;

374 or a heavy chain variable domain amino acid sequence of SEQ ID NO;

88 and a light chain variable domain amino acid sequence of SEQ ID NO;

378, or an antigen-binding portion thereof, to form a complex between said second monoclonal antibody, or antigen-binding portion thereof, and said complex formed in step (i), and

(iii) detecting said complex formed in step (ii).

2. The method of claim 1, wherein said first and/or second monoclonal antibody, or said antigen-binding portion thereof, is detectably labeled with a label, and wherein said detecting said complex in step (iii) comprises detecting said label.

3. The method of claim 1, wherein said second monoclonal antibody comprises:

a heavy chain variable domain amino acid sequence of SEQ ID NO;

20 and a light chain variable domain amino acid sequence of SEQ ID NO;

374.

4. The method of claim 2, wherein said label is a fluorescent label or acridinium label.

5. The method of claim 1, wherein said first monoclonal antibody is bound to a solid phase.

11. The method of claim 1, wherein said second monoclonal antibody comprises a heavy chain variable domain amino acid sequence of SEQ ID NO:

88 and a light chain variable domain amino acid sequence of SEQ ID NO;

378.

6. A method for the detection of HCV protein in a test sample comprising:

(i) contacting a test sample suspected of containing HCV with a first monoclonal antibody comprising a heavy chain variable domain amino acid sequence of SEQ ID NO;

20 and a light chain variable domain amino acid sequence of SEQ ID NO;

374 or a heavy chain variable domain amino acid sequence of SEQ ID NO;

88 and a light chain variable domain amino acid sequence of SEQ ID NO;

378, or an antigen-binding portion thereof, to form a complex between said first monoclonal antibody, or said antigen-binding portion thereof, and said HCV core protein located within said test sample;

(ii) contacting said complex formed in step (i) with a second monoclonal antibody, or antigen-binding portion thereof, which specifically binds to an epitope in the DNA binding domain of HCV core protein to form a complex between said second monoclonal antibody, or said antigen-binding portion thereof, and said complex formed in step (i), and

(iii) detecting said complex formed in step (ii).

7. The method of claim 6, wherein said first and/or second monoclonal antibody, or said antigen-binding portion thereof, is detectably labeled with a label, and wherein said detecting said complex in step (iii) comprises detecting said label.

8. The method of claim 6, wherein said first monoclonal antibody comprises:

a monoclonal antibody comprising a heavy chain variable domain amino acid sequence of SEQ ID NO;

20 and a light chain variable domain amino acid sequence of SEQ ID NO;

374.

9. The method of claim 7, wherein said label is a fluorescent label or an acridinium label.

10. The method of claim 6, wherein said first monoclonal antibody is bound to a solid phase.

12. The method of claim 6, wherein said first monoclonal antibody comprises a heavy chain variable domain amino acid sequence of SEQ ID NO:

88 and a light chain variable domain amino acid sequence of SEQ ID NO;

378.